Thioflavin S and Congo Red inhibit adhesion of Cryptosporidium parvum to HCT-8 cells resulting in a decreased endoplasmic stress response – Part I

Greetings! My name is Dustin Lee and I’m an undergraduate at Pace University (NYC). You may/may not have already read my previous blog posts, but we, my mentor Dr. Cho Chan, and I are continuing our research on the parasite known as Cryptosporidium parvum, but on a larger and different scale. We are using the data acquired from the earlier results and basing our current project off the new information. Basically, we learned that specific concentrations of amyloid-perturbing dyes Thioflavin S and Congo Red reduces adhesion of C.parvum to the host intestinal adenocarcinoma HCT-8 cell line. This is fantastic because adhesion is an essential stage for infecting host cells. With that being said, our current project is titled “Thioflavin S and Congo Red inhibit adhesion of Cryptosporidium parvum to HCT-8 cells resulting in a decreased endoplasmic stress response.” It’s crucial to know some background about the C.parvum parasite; hence I’m going to provide a little lowdown about the parasite. C.parvum is an intracellular yet extracytoplasmic protozoan parasite that induces cryptosporidiosis. Cryptosporidiosis, or crypto, is a gastrointestinal illness that causes acute, watery diarrhea for people with a competent immune system. Although for those who are immunocompromised, the symptoms are usually more fatal. Additionally, other symptoms include dehydration, fever, abdominal pain, and nausea/vomiting, and can last for a couple weeks. Crypto is a widespread waterborne disease found around the globe and is spread through the fecal-oral route, usually from contaminated water.

Dr. Chan and I are studying Cryptosporidium parvum, primarily the role of its amyloid-forming sequences in adhesion proteins. With the preceding data using bioinformatics software programs to confirm the presence of amyloids in C.parvum adhesion proteins, we have verified the amyloid positive sequences in the adhesion proteins. Also using the results from the prior project, we now know that these amyloids mediate adhesion. Our conjecture was that if amyloid formation in the sporozoites’ adhesion proteins were inhibited, there would be a decreased infection of host HCT-8 cells, and we were correct. Now, we need to ascertain the most effective concentration that inhibits amyloid formation by running protein assays with different dye concentrations. It is important to test the efficacy of different dye concentrations because we want to reduce the endoplasmic stress response that is brought about from infection of HCT-8 cells by C.parvum. When C.parvum infects the host HCT-8 cells, an endoplasmic stress response is triggered and putrescine levels in HCT-8 increase about 2.5-fold. We want to decrease the host HCT-8 cells’ stress response by using the amyloid-perturbing dyes, which is essentially the basis of our project.

I’m very thankful to the Undergraduate Student-Faculty Research Program offered at Pace University. I’ve been lucky enough to be chosen/accepted twice as a recognized student researcher, and for that I am grateful. I still highly recommend going under the tutelage of a mentor in your field of interest, or even in a subject that generally peaks your interest. It will indeed teach you something that cannot be learned in a classroom.

 

Signing off,

Dustin Lee